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Objective:To investigate the screening values of immunocytochemical P16/Ki-67 double staining, P16 INK4α single staining and high-risk human papillomavirus (HR-HPV) testing for high-grade cervical lesions. Methods:The clinical data of 622 patients who underwent cervical thin-layer liquid-based cytology (TCT) and HR-HPV testing in General Hospital of Taiyuan Iron and Steel (Group) Co., Ltd. from March 2019 to July 2021 were retrospectively analyzed. The remaining cytological specimens were detected by P16/Ki-67 double staining and P16 INK4α single staining. Among them, 334 patients with TCT results suggesting atypical squamous cell of undetermined significance (ASCUS) and above and HPV-positive underwent colposcopy pathological biopsy. Using pathological results as reference, the positive predictive value, sensitivity, specificity and accuracy of P16/Ki-67 double staining, P16 INK4α single staining and HR-HPV testing for screening of high-grade squamous intraepithelial neoplasia (HSIL) and cervical cancer were compared. Results:Taking the results of histopathology as references, combined with the results of TCT, 31 of 622 patients were HSIL, of which 22 (71.0%) were positive for P16/Ki-67 double staining, 23 (74.2%) were positive for P16 INK4α single staining, and 25 (80.6%) were positive for HR-HPV testing; 4 cases were cervical cancer, and the positive rates of the three detection methods were all 100.0% (4/4). Among 622 patients, the positive rates of P16/Ki-67 double staining, P16 INK4α single staining and HR-HPV testing for screening of HSIL and cervical cancer were 13.99% (87/622), 25.40% (158/622) and 21.38% (133/622); the positive predictive values were 29.89%, 17.09% and 21.08%; the accuracies were 91.19%, 78.94% and 83.28%; the specificities were 89.77%, 77.98% and 82.46%; the sensitivities were 74.29%, 77.14% and 82.86%. The positive rate, positive predictive value, specificity and accuracy of P16/Ki-67 double staining were higher than those of P16 INK4α single staining and HR-HPV testing, and the differences were statistically significant ( z values were -5.062 and -3.418, 2.328 and 2.450, 5.436 and 3.570, 6.043 and 4.161, all P < 0.05); the sensitivity of HR-HPV testing was higher than that of P16/Ki-67 double staining and P16 INK4α single staining, but the differences were not statistically significant ( z values were -0.890 and 1.017, both P > 0.05). Conclusions:HR-HPV testing is more suitable for primary cervical lesion screening; P16/Ki-67 double staining can be used as a potential combined cell screening tool or an effective triage tool; P16 INK4α single staining has certain limitations.
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Objective:To explore the detection capability of p16/Ki-67 double staining technique in women with various abnormal thinprep cytologic test (TCT) results and its diagnostic value for cervical intraepithelial neoplasia Ⅱ+ grade (CIN2+).Methods:A total of 225 women with abnormal TCT results, i.e. the atypical squamous cells of undetermined significance(ASC-US), in the Tianjin Central Hospital of Gynecology Obstetrics, Nankai University Affiliated Maternity Hospital from December 2018 to December 2019 were enrolled. p16/Ki-67 double staining were detected and compared with the high risk human papillomavirus (HR-HPV) and pathological results.Results:The positive rates of p16/Ki-67 double staining increased with cytologic and pathologic categories. For diagnosis of CIN2+, p16/Ki-67double staining (90.1%) was less sensitive than HR-HPV testing (98.2%)( P<0.05), but the specificity of p16/Ki-67 double staining (58.8%) was significantly higher than HR-HPV(21.6%) ( P<0.001). Conclusions:Compared with HR-HPV, p16/Ki-67 double staining has better effect on diagnosing CIN2+. p16/Ki-67 double staining can be considered as triaging method for management of ASC-US and LSIL patients, significantly reduce the colposcopy referral rate (nearly 50%), which has high clinical application value.
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Objective To investigate the diagnostic value of DNA ploidy analysis combined with immunocytochemistry p16/ki-67 double staining in cervical high grade squamous intraepithelial neoplasia(HSIL) and cervical squamous cell carcinoma(SCC).Methods A total of 73 cases of cytological tests were randomly collected.Among them,53 cases were small DNA ploidy abnormal cells and 20 cases were DNA ploidy negative.The p16/Ki-67 results were detected by immunocytochemistry double staining.With the pathological results as the golden standard,the diagnostic values of DNA ploidy analysis and DNA ploidy analysis combined with p16/Ki-67 double staining in HSIL + was contrastively analyzed by pathologic results.Results Among 20 samples of DNA ploidy negative,the p16/Ki-67 double staining results all were negative.The positive predictive value of DNA ploidy analysis for HSIL + was 34.62％.The sensitivity of DNA ploidy analysis combined with p16/Ki-67 double staining for HSIL + was 84.62％,and its specificity was 92.31％,the positive predictive value was 78.57％ and the negative predictive value was 94.74％,which were significantly higher than those of DNA ploidy analysis(P＜0.05).Conclusion p16/Ki-67 double staining can significantly im prove the prediction value of HSIL.The DNA ploidy analysis combined with p16/Ki-67 double staining is an effective method for predicting HSIL +,which is suitable for the implementation in the areas with lack of medical resources.
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BACKGROUND: Melanomas need to be differentiated from benign melanocytic lesions during diagnosis. However, it is difficult to differentiate them using histopathology alone, since both neoplasms have broad morphological spectrums and subtle differentiating features. OBJECTIVE: To evaluate the usefulness of Ki-67/Melan-A double staining for differentiating melanoma from benign melanocytic nevi. METHODS: We selected 20 cases of intradermal nevi, 20 cases of compound nevi, 5 cases of dysplastic nevi, and 25 cases of melanoma from clinicopathologically proven cases reviewed by the Department of Dermatology at our medical center. Ki-67/Melan-A double staining was performed, and the Melan-A verified Ki-67 index (Ki-67-M index) and Ki-67 index were measured. The immunopositivity was measured in the deepest third of the lesions. RESULTS: The Ki-67-M index of intradermal nevi, compound nevi, dysplastic nevi, and melanoma were 0.4+/-0.9%, 1.0+/-1.1%, 4.3+/-1.7%, and 24.1+/-10.9%, respectively. The best Ki-67/Melan-A cut-off point to distinguish melanomas from benign melanocytic nevi was 5%; the sensitivity and specificity were 100% and 97.7%, respectively. Immunopositivity in the deepest third of the intradermal nevi, compound nevi, and melanoma, were 10.5%, 20%, and 100%, respectively; the sensitivity and specificity for diagnosing melanoma were 100% and 84.6%, respectively. The sensitivity and specificity of combined Ki-67-M and immunopositivity in the deepest third for diagnosing melanoma were 100% and 97.7%, respectively. CONCLUSION: The Ki-67-M index and immunopositivity in the deepest third of melanoma were significantly higher than that of benign melanocytic nevi. Therefore, Ki-67/Melan-A double staining is a potentially valuable diagnostic tool for differentiating melanoma from benign melanocytic nevi.
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Dermatology , Diagnosis , Dysplastic Nevus Syndrome , MART-1 Antigen , Melanoma , Nevus , Nevus, Intradermal , Nevus, Pigmented , Sensitivity and SpecificityABSTRACT
PURPOSE: To assess the clinical outcomes in idiopathic epiretinal membrane (ERM) patients after vitrectomy and ERM removal with or without additional indocyanine green (ICG)-assisted internal limiting membrane (ILM) peeling. METHODS: The medical records of 43 patients with an idiopathic ERM that underwent vitrectomy and ERM removal between July 2007 and April 2010 were reviewed. The patients were divided into two groups: triamcinolone-assisted simple ERM peeling only (group A, n = 23) and triamcinolone-assisted ERM peeling followed by ICG staining and peeling of the remaining internal ILM (group B, n = 20). RESULTS: No difference was found between the two groups in terms of visual acuity, macular thickness, P1 amplitude or implicit time on multifocal-electroretinogram (mfERG) at six and 12 months postoperatively. In group B, ICG staining after ERM peeling demonstrated that the ILM had been removed together with the ERM in 12 eyes (60%), and all 12 eyes showed punctate retinal hemorrhages during ERM peeling. There was no recurrence of an ERM in either group. CONCLUSIONS: Additional procedures involving ICG staining and ILM peeling during ERM surgery do not appear to have an additive effect on the clinical outcomes in terms of visual acuity, retinal function based on mfERG, or recurrence rate.
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Aged , Female , Humans , Male , Middle Aged , Coloring Agents , Epiretinal Membrane/surgery , Follow-Up Studies , Indocyanine Green , Postoperative Complications/diagnosis , Retinal Hemorrhage/diagnosis , Retrospective Studies , Treatment Outcome , Visual Acuity , VitrectomyABSTRACT
Objective To investigate the correlation of PARP with the metastasis of colorectal carcinoma.Methods Poly(ADP-ribose) polymerase activity was tested by detecting poly-ADP ribose.Immunohistochemical SP was used to detect the expression of PAR,P-selectin and ICAM-1 in colorectal carcinoma and control colorectal mucosa.The co-expression of PAR with P-selectin and ICAM-1 was investigated by immunofluorescence staining in colorectal carcinoma and control colorectal mucosa.Results The expression of PAR,P-selectin and ICAM-1 was significantly higher in colorectal carcinoma than that in the control colorectal mucosa((P
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Amygdala (AM) plays crucial roles in emotional learning, memory and behavior. These functions of AM are carried out by three main subnuclei (lateral nucleus, basolateral nucleus and central nucleus) in AM and closely related with a transcription factor, cAMP- responsive element binding protein (CREB) in the neurons of the AM. CREB can be phosphorylated (pCREB) in many kinds of neuronal processes to regulate the synthesis of proteins for the formation of memory processes. In order to identify what neuronal types express pCREB and how the pCREB levels changed at different time intervals after an emotional stress stimulation, the present study is designed to investigate pCREB-, glutamate (Glu)- and parvalbumin (PV)- immunoreactive (IR) profiles in AM and the levels of pCREB in AM after a stress of forced swimming (FS). The results showed that the pCREB expressed in the Glu-IR neurons but not in the PV-IR neurons, and the expression level of the pCREB increased dramatically after the stress. The present results suggested that pCREB modulates the emotional processes through the Glu-IR neurons and that the pCREB greatly upregulated to response to the emotional stimuli.
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@#ObjectiveTo observe the expression of vascular endothlial growth factor(VEGF) and throbospondin-1(TSP-1) in different grade bladder cancer.MethodsSpecimens of 70 cases of bladder transitional cell cancer including Grade Ⅰ 28 cases,Grade Ⅱ 27 cases,Grade Ⅲ 15 cases(according to WHO pathological grade) were stained with immunohistochemistry method.The expressing values of VEGF and TSP-1 were calculated by mean vascular density(MVD).All data was analyzed with SPSS 10.0,and with expression of VEGF and TSP-1 was detected with immunohistochemisrty double staining.Finally,the result of expression was observed by laser scan with focus microscope.ResultsThere was a positive correlation between the cancer grade and the expression of VEGF,and the expression of Grade Ⅰ was significant difference with that of Grade Ⅱ(P<0.01),but there was no difference between Grade Ⅱ and Grade Ⅲ(P>0.1).Expression of TSP-1 decreased with grade increasing but not showing a negative correlation,while,Grade Ⅰ was significant difference with that of Grade Ⅱ and Grade Ⅲ(P<0.01),Grade Ⅱ was not differcnt with Grade Ⅲ(P>0.1).In 28 Grade Ⅰ cases,values of MVD of VEGF and TSP-1 had no correlation(rs)=0.167,P>0.1).Double immunostaining detecting with laser scan with focus microcope indicate showed that VEGF and TSP-1 appeared in same bladder cancer specimen.ConclusionExpression of VEGF is gradually increased from Grade Ⅰ to Grade Ⅲ,and VEGF is a promoting factor of angiogenesic bladder cancer.Expression of TSP-1 is the strongest in Grade Ⅰ cases,and TSP-1 is an inhibiting angiogenesic factor,which inhibiting function only in early stage.Immunofluorescent staining can only provide evidence of together expression of VEGF and TSP-1,cannot evaluate the degree of expression.
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Objective To study the human lymphangiogenesis in the early stage of embryo and the expression of forkhead box C2(FOXC2) in human lymphangiogenesis.Methods Lymphatic vessel endothelial haluronic acid receptor-1(LYVE-1) was used as the special marker of lymphatic vessels.Immunohistochemical and double immunofluorescence staining were used to detect the expressions of FOXC2 and LYVE-1 in lymphatic vessels of 85 human embryos of 5 to 11 weeks pregnancy and analyze the features of lymphatic vessel genesis and development.Results LYVE-1 was initially detected in lymphatic vessels in 7-week human embryos.The lymphatic vessel endothelial cells presented brown staining for LYVE-1 in jugular and thoracic region.LYVE-1 was also found to express in human embryonic mesentery lymphatic vessels on the 70th day of pregnancy.The expression of FOXC2 in human embryo was detected prior to that of LYVE-1.At the beginning of the 6th week of pregnancy,FOXC2 was obviously seen in human embryonic mesoderm mesenchyme.FOXC2 expressed not only in human embryonic lymphatic vessels,but also in the vertebral body,cardiovascular wall and other tissues.The expressions of FOXC2 and LYVE-1 were still seen in human embryonic lymphatic vessel endothelial cells after 80 days of pregnancy.Conclusion Lymphangiogenesis of human embryo begins between the 7th and 8th weeks of pregnancy.FOXC2 and LYVE-1 could have some relation with the human embryonic lymphatic vessel genesis and development.
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This experiment developed the methodology of double staining for senescence associated-beta-galactosidase (SA-beta-gal) activity and keratin 10 (K10) or involucrin. To prove the usefulness of the double staining, the author investigated the relationship between senescence and differentiation in monolayer and organotypic cultured keratinocytes. The results were as follows: K10 and involcrin together with SA-beta-gal were doubly stained in most of monolayer cultured keratinocyte. This fact indicated that the senescence and differentiation had simultaneously occurred in the same keratinocyte. In spite of the advantages to preserving structures, the paraffin specimen was not suitable for double staining because of the limitation of SA-beta-gal reactivity. Although the cryosectioned specimen did not have the morphology as good as the paraffin specimen, it was suitable for double staining due to the goodness of SA-beta-gal reactivity. Double staining well reflected the disturbances of senescence and differentiation which could be caused by deranged organizations of the organotypic cultured skin. The organotypic cultured skin which showed deranged organizations such as stratified basal layer, no typical cell features in each epdermal layer, and wide intercellular spaces had SA-beta-gal activity in epidermis and K10 or involucrin reaction in basal cell. But the skin which showed well arranged organizations resembling in vivo skin had no SA-beta-gal activity and no K10 or involucrin reaction in basal cells. In conclusion, it might be suggested that the double staining for SA-beta-gal activity and K10 or involucrin could be used for detecting the extent of senescence and differentiation in the same cell.
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Aging , Epidermis , Extracellular Space , Keratin-10 , Keratinocytes , Paraffin , SkinABSTRACT
Objective:To discuss the role of Fas/FasL-mediated colonic epithelial apoptosis in the pathogenesis of ulcerative colitis(UC).Methods:Expression of Fas,FasL and M30(a new epithelium apoptosis marker) antigen on colonic mucosa of 35 patients with UC and 15 control populations were detected by IHC.Each marker was analyzed by image analysis system.Co-expression of Fas and M30 was observed through serial sections and double staining IHC.Results:The expression of Fas,FasL as well as M30 in UC increased considerably compared with controls (P
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Objective To observe the connections between enkephalin immunoreactive(ENK-ir) terminals and ?-aminobutyric acid immunoreactive(GABA-ir) neurons in the rostral portion of the nucleus tractus solitarious(rNTS) of the rat. Methods Double-immunofluorescent labeling method and the pre-embedding immunohistochemical staining combined with immuno-gold particles labeling technique for electron microscopic detection were used in the present study. Results Under the laser scanning confocal microscope,dense ENK-ir fibers and terminals and some GABA-ir neurons were observed in the rNTS.Close contacts between ENK-ir terminals and GABA-ir cell bodies were also detected.By using the electron microscopic technique,ENK-ir reaction products were found to be mainly localized on the surface of round clear vesicles and dense-cored vesicles in the axon terminals.ENK-ir terminals formed symmetric,which was a dominant pattern,and asymmetric synaptic connections with GABA-ir and immunonegative cell bodies and dendrites.Conclusion The ENK-ir terminals might take part in the transmission and regulation of the taste information in the rNTS through inhibiting,exciting the GABAergic neurons or inhibiting directly the activity of neurons within the rNTS.
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By using 5'-nucleotidase-alkaline phosphatase (5'-Nase-AlPase) double staining method, the fine distribution of the lymph vessels in the stomach wall of the rabbit, guinea pig and rat has been observed. Under light microscope lymph capillaries showed strongly 5'-Nase positive reaction with brown or dark brown staining. However blood capillaries revealed significantly A1Pase activity with blue staining. The lymph capillaries were seen in all layers of the stomach wall in three species, but the lymph vessels were only shown in the submucosa, muscularis and serosa. In the mucosa lymph capillaries could be found in the deep layer of the lamina propria. In the submucosa there were lymph vessels and capillaries. In the muscularis lymph capillaries and vessels not only could be found between oblique and circular muscle or between circular and longitudinal muscle, but also among muscle fiber bundles of each layer. In the serosa there were larger lymph vessels situated near the muscularis. The observation on semithin and ultrathin sections also shown the similarity in the lymph vessels distribution. There was no difference in the distribution of lymph vessels in the stomach wall between rabbit, guinea pig and rat.